Cell Signaling Technology

Product Pathways - Motif Antibodies

Phospho-(Ser) 14-3-3 Binding Motif Antibody #9601

14-3-3   general   KXXSXP   motif   PBS   phospho serine   phospho-serine   phosphoserine   RXXSXP   substrate  

No. Size Price
9601S 100 µl ( 10 western blots ) ¥3,900.00 现货查询 购买询价
9601 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Rat,All Species Expected, Endogenous Rabbit
IP 1:50
IHC-P 1:50
E-P 1:1000

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, IHC-P=Immunohistochemistry (Paraffin), E-P=Peptide ELISA (DELFIA),

Specificity / Sensitivity

Phospho-(Ser) 14-3-3 Binding Motif Antibody binds peptides and proteins containing a motif composed of phospho-Ser with proline at the +2 position and arginine or lysine at the -3 position. Antibody binding is phospho-specific and largely independent of other surrounding amino acids. The antibody weakly cross-reacts with sequences containing phospho-Thr instead of phospho-Ser in this motif, and with sequences containing phospho-Ser surrounded by Phe at the +1 position and Arg/Lys at the -3 position. No cross-reactivity is observed with corresponding nonphosphorylated sequences or with other phospho-Thr/Ser/Tyr-containing motifs. By ELISA this antibody recognizes a wide range of peptides containing the 14-3-3 binding motif, and by 2D gel Western blot analysis it recognizes a large number of presumptive 14-3-3 binding proteins. (U.S. Patent No's.: 6,441,140; 6,982,318; 7,259,022; 7,344,714; U.S.S.N. 11,484,485; and all foreign equivalents.)

Phospho-(Ser) 14-3-3 Binding Motif Antibody 兔多抗结合+2位脯氨酸和-3位精氨酸或赖氨酸的磷酸化丝氨酸肽段或蛋白。该抗体是包含磷酸化特异性的,并在很大程度上与周围氨基酸序列无关。该抗体与基序中包含磷酸化苏氨酸而非磷酸化丝氨酸的序列有弱相互作用,并与包含+1位有苯丙氨酸或-3位有精氨酸/赖氨酸的磷酸化丝氨酸肽段有弱相互作用。没有发现其能与相应的非磷酸化的序列或其它基序序列中的磷酸化苏氨酸/丝氨酸/酪氨酸序列发生交叉反应。在ELISA中,其可以识别多种包含14-3-3结合基序的肽段,在二维凝胶Western Blot中,其可以识别很多预测的14-3-3结合蛋白。(美国专利号:6,441,140、6,982,318、7,259,022、7,344,714;U.S.S.N. 11,484,485;及所有国外相应专利)

Source / Purification

Polyclonal antibodies are produced by immunizing animals with synthetic phospho-(Ser) 14-3-3-binding-motif containing peptides . Antibodies are purified by protein A and peptide affinity chromatography.

该多克隆抗体用人工合成的包含磷酸化丝氨酸14-3-3结合基序的肽段免疫动物制备。该抗体使用蛋白A和蛋白亲和层析纯化。

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, showing staining of proteins containing phosphorylated 14-3-3 binding motifs, using Phospho-(Ser) 14-3-3 Binding Motif Antibody.

对石蜡包埋的人乳腺癌使用Phospho-(Ser) 14-3-3 Binding Motif Antibody进行免疫组化分析,显示包含磷酸化14-3-3结合基序的蛋白染色。

Western Blotting

Western Blotting

Western blot analysis of extracts from 293 cells transfected with Bad or Bad Ser112 and/or Ser136 mutant fusion proteins, using Phospho-(Ser) 14-3-3 Binding Motif Antibody (top), Phospho-Bad (Ser136) Antibody #9295 (middle) or Phospho-Bad (Ser112) Antibody #9291 (bottom).

对293细胞(转染Bad或Bad Ser112和/或Ser136突变融合蛋白),使用Phospho-(Ser) 14-3-3 Binding Motif Antibody(上图),Phospho-Bad (Ser136) Antibody #9295(中图)或Phospho-Bad (Ser112) Antibody #9291(下图)进行Western blot分析。

Western Blotting

Western Blotting

Western blot analysis of extracts from Jurkat cells, untreated or calyculin A-treated (0.1 µM for 30 minutes), using Phospho-(Ser) 14-3-3 Binding Motif Antibody. Proteins were separated by 2-D electrophoresis prior to blotting.

对未处理或0.1uM calyculin A裂解前处理30分钟的Jurkat细胞抽提液使用Phospho-(Ser) 14-3-3 Binding Motif Antibody进行Western blot分析。蛋白质在印记前在二维电泳分离。

ELISA-Peptide

ELISA-Peptide

Phospho-(Ser) 14-3-3 Binding Motif Antibody ELISA Assay: Signal-to-noise ratio of phospho- versus nonphospho-14-3-3 binding motif peptides. (S* denotes phosphorylated serine.)

Phospho-(Ser) 14-3-3 Binding Motif Antibody ELISA分析:磷酸化和非磷酸化14-3-3结合基序的信噪比。(S* 代表磷酸化的丝氨酸)

Background

The 14-3-3 proteins are a highly conserved family of proteins involved in the regulation of cell survival, apoptosis, proliferation and checkpoint control (1-5). Biological regulation by 14-3-3 is mediated through phosphorylation-dependent protein-protein interactions (6). Two different phospho-Ser-containing motifs are found within nearly all known 14-3-3 binding proteins (7). Motif 1 (Arg/Lys and Ser at positions -3 and -2, phospho-Ser at position 0, and Pro at position +2) is found in critical regulatory proteins including Bad, cdc25C, FKHRL1, PKC and c-Raf (5,7). Phospho-(Ser) 14-3-3 Binding Motif Polyclonal and (4E2) Monoclonal Antibodies provide powerful tools for the discovery and characterization of potential 14-3-3 binding proteins containing this motif and for high throughput drug discovery.

14-3-3蛋白是参与调控细胞生存、凋亡、增殖和检查点控制的一组高度保守的蛋白家族(1-5)。14-3-3的生物调控由磷酸化依赖的蛋白-蛋白相互作用介导(6)。两种不同的包含磷酸化丝氨酸的基序在几乎所有的14-3-3结合蛋白中发现(7)。基序1(精氨酸/赖氨酸和丝氨酸在-3和-2位,磷酸化丝氨酸在0位,脯氨酸在+2位)在很多重要的调节蛋白包括Bad、cdc25C、FKHRL1、PKC和cRaf中发现(5-7)。Phospho-(Ser) 14-3-3 Binding Motif Polyclonal and (4E2) Monoclonal Antibody(结合基序多克隆和单克隆抗体)提供了发现和鉴定潜在的含此基序的14-3-3结合蛋白、及高通量药物发现的有力工具。

  1. Aitken, A. (1995) Trends Biochem Sci 20, 95-7.
  2. Zha, J. et al. (1996) Cell 87, 619-28.
  3. Piwnica-Worms, H. (1999) Nature 401, 535, 537.
  4. Tzivion, G. et al. (1998) Nature 394, 88-92.
  5. Xing, H. et al. (2000) EMBO J 19, 349-58.
  6. Muslin, A.J. et al. (1996) Cell 84, 889-97.
  7. Yaffe, M.B. et al. (1997) Cell 91, 961-71.

Application References

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For Research Use Only. Not For Use In Diagnostic Procedures.

Use of Cell Signaling Technology (CST) Motif Antibodies within certain methods (e.g., U.S. Patents No. 7,198,896 and 7,300,753) may require a license from CST. For information regarding academic licensing terms please have your technology transfer office contact CST Legal Department at CST_ip@cellsignal.com. For information regarding commercial licensing terms please contact CST Pharma Services Department at ptmscan@cellsignal.com.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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